Cyclic n,n&#39;-diarylthioureas and n,n&#39;-diarylureas - androgen receptor  antagonists, anticancer  agent, method for preparation and use thereof

ABSTRACT

The present invention relates to novel cyclic N,N′-diarylureas and N,N′-diarylthioureas—androgen receptor antagonists, anti-cancer agent, pharmaceutical composition, medicament, and method for treatment of cancerous diseases, among them prostate cancer. 
     Cyclic N,N′-diarylthioureas or N,N′-diarylureas of the general formula 1, their optical (R)- and (S)-isomers and pharmaceutically acceptable salts thereof exhibiting properties of androgen receptor antagonists have been proposed, 
     
       
         
         
             
             
         
       
     
     wherein: X represents oxygen or sulfur;
     m=0 or 1;   R1 represents C 1 -C 3 alkyl;   R2 and R3 represent hydrogen; or   R2 and R3 together with C-atom they are attached to form C═O group; R4 and R5 represent hydrogen; or   R4 represents hydrogen, R5 represents methyl; or   R4 represents methyl, R5 represents CH 2 R6 group in which R6 represents C 1 -C 3  alkoxycarbonyl, carboxyl, hydroxyl group optionally substituted with methyl or benzyl; or   R4 and R5 together with C-atom they are attached to form 5- or 6-membered heterocycle comprising at least one oxygen atom or nitrogen atom optionally substituted with methyl; or   R4 and R5 together with C-atom they are attached to form NH group.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a National stage of International applicationPCT/RU2011/000476 filed Jul. 1, 2011, which claims benefit of foreignpriority to the Russian Federation application RU 2010130618 of Jul. 22,2010. The priority applications are hereby incorporated by reference intheir entirety.

FIELD OF THE INVENTION

The invention relates to novel cyclic N,N′-diarylthioureas andN,N′-diarylureas—androgen receptor antagonists, anticancer agent,pharmaceutical composition, medicament and method for treatment ofcancer including prostate cancer.

PRIOR ART

There are known androgen receptor antagonists which are-1,3-diaryl-5,5-dimethyl-2-thioxoimidazolidin-4-ones I,5,7-diaryl-6-thioxo-5,7-diazaspiro[3,4]octan-8-ones II and1,3-diaryl-2-thioxo-1,3-diazaspiro[4,4]nonan-4-ones III exhibitinganticancer activity [WO2006124118, WO2007127010]. Amongst thesecompounds the most promoted is4-[3-[4-cyano-3-(trifluoromethyl)phenyl]-5,5-dimethyl-4-oxo-2-thioxoimidazolidin-1-yl]-2-fluoro-N-methylbenzamideMDV3100 (androgen receptor antagonist with IC₅₀=36 nM), which is now inthe III phase of clinical trials as medicament for prostate cancertreatment [Drug Data Rep., 2009, 31(6), 609].

Searching for highly effective anticancer medicaments exhibitingenhanced activity and reduced toxicity, is still one of the maindirections in the development of novel pharmacological remedies forcancer treatment, including prostate cancer. In this context thedevelopment of novel anticancer active agents, pharmaceuticalcompositions and medicaments, and also methods for their preparation anduse is of essential importance.

DISCLOSURE OF THE INVENTION

In the context of the invention, the terms are generally defined asfollows:

“Azaheterocycle” means an aromatic or non aromatic mono- or poly-cyclicsystem, comprising at least one nitrogen atom in the cycle.Azaheterocycle may have one or more “cyclic system” substituents.

“Active component” (drug-substance) means a physiologically activecompound of synthetic or other origin (biotechnological, vegetable,animal, microbe and so on), exhibiting pharmacological activity andbeing an active component of pharmaceutical composition, employing inproduction and preparation of medicaments.

“Alkyl” means an aliphatic hydrocarbon straight or branched chain with1-12 carbon atoms. Branched means an alkyl chain with one or more “loweralkyl” substituents. Alkyl group may have one or more substituents ofthe same or different structure (“alkyl substituent”) including halogen,alkenyloxy, cycloalkyl, aryl, heteroaryl, heterocyclyl, aroyl, cyano,hydroxy, alkoxy, carboxy, alkynyloxy, aralkoxy, aryloxy,aryloxycarbonyl, alkylthio, heteroarylthio, aralkylthio, arylsulfonyl,alkylsulfonylheteroaralkyloxy and so on.

“Antagonists” mean ligands which being bound to definite receptors notcause active cellular responses. Antagonists prevent linkage betweenagonists and receptors and by that block specific receptor signaltransmission.

“Aryl” means aromatic mono- or poly-cyclic system with 6-14 carbonatoms, predominantly 6-10 carbon atoms. Aryl may have one or more“cyclic system substituents” of the same or different structure. Phenyl,substituted phenyl, naphthyl, or substituted naphthyl are therepresentatives of aryl groups. Aryl could be annelated with nonaromaticcyclic system or heterocycle.

“Heterocyclyl” means aromatic or saturated mono- or polycyclic systemwith 3-10 carbon atoms, preferably from 5 to 6, wherein one or morecarbon atoms are substituted by one or more heteroatoms, such as N, S orO. Prefix “aza”, “oxa” or “thia” before “heterocyclyl” means that N, Oor S atoms are introduced in the cycle, respectively. Heterocyclyl mayhave one or more “cyclic system sustituents” of the same or differentstructure. N— and S— atoms of heterocyclyl cycle could be oxidized toN-oxide, S-oxide or S-dioxide. Piperidinyl, pyrrolidinyl, piperazinyl,morpholinyl, thiomorpholinyl, thiazolidinyl, 1,4-dioxane-2-yl,tetrahydrofuranyl, tetrahydrothiophenyl and others are examples ofheterocyclyl.

“Hydrate” means stoichiometric or nonstoichiometric compositions ofcompounds or their salts with water.

“Substituent” means a chemical radical attached to scaffold (fragment),for example, “alkyl substituent”, “amino group substituent”, “carbamoylsubstituent”, and “cyclic system substituent”, the meanings of which aredefined in this section.

“Medicament”—is a compound (or mixture of compounds in the form ofpharmaceutical composition) in the form of tablets, capsules,injections, ointments and other ready forms intended for restoration,improvement or modification of physiological functions at humans andanimals, and also for treatment and prophylaxis of diseases,diagnostics, anesthesia, contraception, cosmetology and others.

“Lower alkyl” means a straight or branched alkyl with 1-4 carbon atoms.

“Pharmaceutical composition” means a composition comprising a compoundof general formula 1 and at least one of components selected from thegroup consisting of pharmaceutically acceptable and pharmacologicalycompatible fillers, solvents, diluents, auxiliary, distributing andsensing agents, delivery agents, such as preservatives, stabilizers,disintegrators, moisteners, emulsifiers, suspending agents, thickeners,sweeteners, flavouring agents, aromatizing agents, antibacterial agents,fungicides, lubricants, and prolonged delivery controllers, the choiceand suitable proportions of which depend on the nature and way ofadministration and dosage. Examples of suitable suspending agents areethoxylated isostearyl alcohol, polyoxyethene, sorbitol and sorbitolether, microcrystalline cellulose, aluminum metahydroxide, bentonite,agar-agar and tragacant and their mixtures as well. Protection againstthe action of microorganisms can be provided by various antibacterialand antifungal agents, such as, for example, parabens, chlorobutanole,sorbic acid, and similar compounds. Composition may also containisotonic agents, such as, for example, sugar, sodium chloride, andsimilar compounds. Prolonged effect of composition may be achieved byagents slowing down absorption of active ingredient, for example,aluminum monostearate and gelatine. Examples of suitable carriers,solvents, diluents and delivery agents include water, ethanol,polyalcohols and their mixtures, natural oils (such as olive oil) andorganic esters (such as ethyl oleate) for injections. Examples offillers are lactose, milk-sugar, sodium citrate, calcium carbonate,calcium phosphate and the like. Examples of disintegrators anddistributors are starch, alginic acid and its salts, and silicates.Examples of suitable lubricants are magnesium stearate, sodium laurylsulfate, talc and polyethylene glycol of high molecular weight.Pharmaceutical composition for peroral, sublingval, transdermal,intramuscular, intravenous, subcutaneous, local or rectal administrationof active ingredient, alone or in combination with another activecompound may be administered to humans and animals in standardadministration form, or in mixture with traditional pharmaceuticalcarriers. Suitable standard administration forms include peroral formssuch as tablets, gelatin capsules, pills, powders, granules,chewing-gums and peroral solutions or suspensions, sublingval andtransbuccal administration forms; aerosols; implants; local,transdermal, subcutaneous, intramuscular, intravenous, intranasal orintraocular forms of introduction and rectal administration forms.Pharmaceutical compositions are usually prepared by means of standardprocedures by mixing an active compound with liquid or overgroundedsolid carrier.

“Pharmaceutically acceptable salt” means a relatively nontoxic bothorganic and inorganic salts of acids and bases disclosed in thisinvention. Salts could be prepared in situ in processes of synthesis,isolation or purification of compounds or be prepared specially. Inparticular, bases salts could be prepared starting from purified base ofdisclosed compound and suitable organic or mineral acid. Examples ofsalts prepared in this manner include hydrochlorides, hydrobromides,sulfates, bisulfates, phosphates, nitrates, acetates, oxalates,valeriates, oleates, palmitates, stearates, laurates, borates,benzoates, lactates, p-toluenesulfonates, citrates, maleates, fumarates,succinates, tartrates, methane sulphonates, malonates, salicylates,propionates, ethane sulphonates, benzene sulfonates, sulfamates and thelike (Detailed description of properties of such salts is given in:Berge S. M., et al., “Pharmaceutical Salts” J. Pharm. Sci., 1977, 66:1-19). Salts of disclosed acids may be also prepared by reaction ofpurified acids specifically with suitable base; moreover, metal saltsand amine salts may be synthesized too. Metal salts are salts of sodium,potassium, calcium, barium, magnesium, zink, lithium and aluminum,sodium and potassium salts being preferred. Suitable inorganic basesfrom which metal salts can be prepared are sodium hydroxide, carbonate,bicarbonate and hydride; potassium hydroxide, carbonate and bicarbonate,lithium hydroxide, calcium hydroxide, magnesium hydroxide, zinchydroxide. Organic bases suitable for preparation of disclosed acidsalts are amines and amino acids of sufficient basicity to producestable salt suitable for medical purposes use (in particular, they areto have low toxicity). Such amines include ammonia, methylamine,dimethylamine, trimethylamine, ethylamine, diethylamine, triethylamine,benzylamine, dibenzylamine, dicyclohexylamine, piperazine,ethylpiperidine, tris(hydroxymethyl)aminomethane and the like. Besides,salts can be prepared using some tetraalkylammonium hydroxides, such asholine, tetramethylammonium, tetraethylammonium, and the like.Aminoacids may be selected among main aminoacids—lysine, ornithine andagrinine.

The authors have disclosed novel cyclic N,N′-diarylthioureas andN,N′-diarylureas of the general formula 1, optical (R)- and (S)-isomersand pharmaceutically acceptable salts thereof which are androgenreceptor antagonists:

wherein:

-   X represents oxygen or sulfur;-   m=0 or 1;-   R1 represents C₁-C₃alkyl;-   R2 and R3 represent hydrogen; or-   R2 and R3 together with the C-atom they are attached to form C═O    group;-   R4 and R5 represent hydrogen; or-   R4 represents hydrogen, R5 represents methyl; or-   R4 represents methyl, R5 represents CH₂R6 group in which R6 is C₁-C₃    alkoxycarbonyl; carboxyl; hydroxyl group optionally substituted with    methyl or benzyl; or-   R5 and R4 together with the C-atom they are attached to form 5- or    6-membered heterocycle comprising at least one oxygen atom or    nitrogen atom optionally substituted with methyl; or-   R4 and R5 together with the C-atom they are attached to represent NH    group.

The preferred compounds are N,N′-diarylthioureas and N,N′-diarylureas,their optical (R)- and (S)-isomers and pharmaceutically acceptable saltsof the general formulas 1.2, 1.3 or 1.4:

wherein:

-   X, R1, R2, R3, R4 and R5 have the above meanings

The more preferable compounds are cyclic N,N′-diarylthioureas offormulas 1.2(1), 1.2(2), 1.2.2 and 1.2.3, their optical(R)-isomers—(R)-1.2(2), (R)-1.2.2, (R)-1.2.3 and (S)-isomers—(S)-1.2(2),(S)-1.2.2 and (S)-1.2.3:

wherein:

-   R5 and R4 together with the C-atom they are attached to form 5- or    6-membered heterocycle comprising at least one oxygen atom or    nitrogen atom optionally substituted with methyl,-   R6 has the meaning mentioned above.

The more preferable compounds are also compounds of formulas 1.2.2(1),1.2.2(2), 1.2.2(3), 1.2.3(1), 1.2.3(2) and 1.2.3(3), their optical(R)-isomers—(R)-1.2.2(1), (R)-1.2.2(2), (R)-1.2.2(3), (R)-1.2.3(1), and(S)-isomers—(S)-1.2.2(1), (S)-1.2.2(2), (S)-1.2.2(3), (S)-1.2.3(1), or apharmaceutically acceptable salt thereof,

wherein R6 represents hydroxyl group optionally substituted with methylor benzyl,

The subject of the present invention is a method for preparation ofcompounds of the general formula 1.2 and optical (R)- and (S)-isomersthereof.

1,3-Diarylhydantoines of the general formula 1.2 are prepared byinteraction of isothiocyanate 3.2 with the corresponding4-(cyanomethyl)aminobenzamides 4.1 or (4-carbamoylphenylamino)aceticacids 4.2 according to scheme 1.

Optically active cyclic N,N′-diarylthioureas, (R)-1.2 and (S)-1.2isomers, are prepared either from the corresponding optically active(R)-4.1, (R)-4.2, (S)-4.1 and (S)-4.2 starting materials, or byresolution of racemic mixtures of cyclic N,N′-diarylthioureas 1.2 toenantiomers.

wherein:

-   R1, R4 and R5 have the above meanings.

1,3-Diaryltetrahydropyrimidin-2-ones of the general formula 1.3.1 areprepared by interaction of the corresponding N,N′-diarylureas of thegeneral formula 2 with 1,3-dibromopropane according to scheme 2.

Compounds of the general formula 1.3.2 are prepared by interaction ofisocyanate 3.1 or isothiocyanate 3.2 with the corresponding ethyl13-alaninates of the general formula 5 with subsequent cyclization ofobtained ureas of the general formula 6 according to scheme 3.

1,4-Diaryl[1,2,4]triazolidin-3,5-diones of the general formula 1.4 areprepared by interaction of the corresponding hydrazine 7 with isocyante3.1 and subsequent condensation of the prepared semicarbazide 8 withdiphosgene according to scheme 4.

Novel androgen receptor antagonists are also suitable for investigationof molecular mechanism of inhibition and activation of androgenreceptors.

Novel cyclic N,N′-diarylthioureas and N,N′-diarylureas of the generalformula 1 are androgen receptor antagonists, at that their activityexceeds the activity of known androgen receptor antagonists, publishedin patent application WO2006124118, WO2007127010, and in Drug Data Rep.,2009, 31(6), 609.

Besides, novel antagonist 1.2.2(1) is more than three times as lesstoxical as MDV3100 antagonist, because its maximum tolerated dose (MTD),determined in experiments with male mice of CD1 line is equal to MTD>100mg/kg, whilst MTD for MDV3100 is about ˜30 mg/kg.

The subject of the present invention is novel anticancer agentrepresenting at least one cyclic N,N′-diarylthioureas orN,N′-diarylureas of the general formula 1.

The subject of the present invention is also novel pharmaceuticalcomposition comprising as an active component at least one cyclicN,N′-diarylthiourea or N,N′-diarylurea of the general formula 1, itsoptically active isomer or pharmaceutically acceptable salt exhibitinganticancer activity in effective amount.

The more preferable composition is the pharmaceutical compositionexhibiting activity towards prostate cancer comprising as activecomponent at least one cyclic N,N′-diarylthiourea or N,N′-diarylurea ofthe general formula 1, its optical isomer or pharmaceutically acceptablesalt.

Pharmaceutical compositions may include pharmaceutically acceptableexcipients. Pharmaceutically acceptable excipients mean diluents,auxiliary agents and/or carriers employing in the sphere ofpharmaceutics. According to the invention the pharmaceutical compositionin addition to the cyclic N,N′-diarylthiourea or N,N′-diarylurea of thegeneral formula 1, its optically active isomer or pharmaceuticallyacceptable salt may include other active components, among other thingsexhibiting anti-cancer activity, provided that they do not give rise toundesirable side-effects.

According to the present invention, if it is necessary to use thepharmaceutical composition in clinical practice it can be mixed up withvarious traditional pharmaceutical carries.

According to the present invention the carriers used in pharmaceuticalcompositions represent carriers which are applied in the sphere ofpharmaceutics for preparation of commonly used forms including: bindingagents, greasing agents, disintegrators, solvents, diluents,stabilizers, suspending agents, colorless agents, taste flavors are usedfor peroral forms; antiseptic agents, solubilizers, stabilizers are usedin the forms for injections; base materials, diluents, greasing agents,antiseptic agents are used in local forms.

The purpose of the present invention is also the method for preparationof pharmaceutical compositions.

The object in view is achieved by mixing novel anti-cancer agent with aninert exicipient and/or solvent, the distinctive feature of whichconsists in utilization as anticancer agent, at least, one cyclicN,N′-diarylthiourea or N,N′-diarylurea of the general formula 1, itsoptically active isomer or pharmaceutically acceptable salt.

The subject of the present invention is also a medicament in the form oftablets, sheaths or injections including novel anticancer agent or novelpharmaceutical composition intended for cancer treatment.

The preferred medicament including novel anticancer agent or novelpharmaceutical composition is a medicament intended for treatment ofprostate cancer.

The subject of the present invention is also therapeutic cocktails fortreatment of cancer diseases, among them prostate cancer, including asone of the components novel medicament or novel pharmaceuticalcomposition, comprising as active component at least one cyclicN,N′-diarylthiourea or N,N′-diarylurea of the general formula 1, itsoptically active isomer or pharmaceutically acceptable salt.

Therapeutic cocktails for treatment of prostate cancer, along with themedicament disclosed in the invention, may include other known drugsubstances intended for treatment cancer diseases.

According to the present invention the method for treatment of oncologicdiseases, among them, prostate cancer, at humans and warm-bloodedanimals consists in introduction to human or warm-blooded animal ofnovel medicament, novel pharmaceutical composition or novel therapeuticcocktail.

Medicaments could be administered perorally or parenterally, forexample, intravenously, subcutaneously, intraperitoneally or locally.The clinical dosage of the active component of the general formula 1could be corrected depending on: therapeutic efficiency andbioavailability of the active ingredients in organism, rate of theirexchange and deducing from organism, and also depending on the age, sexand the severity of the patient's symptoms; the daily dosage for adultsfalls within the range of about 10 to about 500 mg of the activeingredient, preferably of about 50 to about 300 mg. Therefore, accordingto the present invention in the process of preparation of a medicamentfrom the pharmaceutical composition as units of dosage it is necessaryto keep in mind the above effective dosage, so that each unit of dosageshould contain of about 10 to about 500 mg of the compound of thegeneral formula 1, preferably 50˜300 mg. In accordance with therecommendation of physician or pharmacist the above dosage can be takenseveral times during the definite time intervals (preferably—from one tosix times).

BEST EMBODIMENT OF THE INVENTION

The invention is illustrated by the following drawings.

FIG. 1. Weight change of male mice at peroral introduction of compound1.2.2(1).

FIG. 2. Weight change of male mice at peroral introduction of compoundMDV3100.

The examples given below describe synthesis of N,N′-diarylthioureas andN,N′-diarylureas and data of their biological investigation, whichillustrate but not limit the scope of the invention.

EXAMPLE 1

Synthesis ofN-methyl-4-{4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl}-2-fluorobenzamide1.2(1). Glycine (80 mg, 1.07 mmol) and K₂CO₃ (207 mg, 1.5 mmol) wereadded to solution of 4-iodo-N-methyl-2-fluorobenzamide (279 mg, 1 mmol)in DMF (3 ml). The reaction mixture was stirred at 140° C. for 18 min.in microwave oven, cooled, diluted with AcOEt (10 ml) and water (10 ml),neutralized with HCl to pH 2-3, organic layer was separated, water layerwas extracted with AcOEt (5×20 ml). The combined extracts were washedwith brine, dried over Na₂SO₄ and evaporated in vacuo. The product wasisolated by colomn chromatography on SiO₂. It gaveN-(4-methylcarbamoyl-2-fluorophenyl)glycine 4.2(1) (R1=CH₃, R4=R5=H,). Asolution of N-(4-methylcarbamoyl-2-fluorophenyl)glycine 4.2(1) (113 mg,0.5 mmol) and 4-isothiocyanato-2-(trifluoromethyl)benzonirile 3.2 (174mg, 1.0 mmol) in DMF (2 ml) was stirred at 90° C. for 12 h. The reactionmixture was evaporated in vacuo, andN-methyl-4-{4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]-imidazolidin-1-yl}-2-fluorobenzamide1.2(1), was isolated by HPLC method, LCMS (M+H)^(|) 437.

EXAMPLE 2

General method for synthesis ofN-methyl-4-{5-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl}-2-fluorobenzamide1.2(2),N-methyl-4-{(S)-5-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl}-2-fluorobenzamide(S)-1.2(2) andN-methyl-4-{(R)-5-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl}-2-fluorobenzamide(R)-1.2(2).

(D,L)-, (D)- or (L)-Alanine (347 mg, 7.8 mmol) and Cs₂CO₃ (2.54 g, 7.8mmol) were added to the solution of N-methyl-2,4-difluorobenzamide (667mg, 3.9 mmol) in DMSO (3 ml). The reaction mixture was stirred in closedvial at 90° C. for 18 h. Cooled mixture was diluted with isopropanol,neutralized with HCl (1.36 ml, 15.6 mmol), filtered, evaporated invacuo, and by HPLC method N-(4-methylcarbamoyl-3-fluorophenyl)alanine4.2(2) (R1=CH₃, R4=H, R5=CH₃),(S)—N-(4-methylcarbamoyl-3-fluorophenyl)alanine (S)-4.2(2) or(R)—N-(4-methylcarbamoyl-3-fluorophenyl)alanine (R)-4.2(2) wereisolated. LCMS (M+H)⁺ 241. ¹H NMR (DMSO-d₆, 400 MHz): 12.66 (br. s, 1H),7.62 (m, 1H), 7.45 (t, J=8.8 Hz, 1H), 6.67 (br. d, J=7.2 Hz, 1H), 6.42(dd, J₁=8.4 Hz, J₂=2.0 Hz, 1H), 6.29 (dd, J₁=14.8 Hz, J₂=2.0 Hz, 1H),4.03 (m, 1H), 2.73 (d, J=4.4 Hz, 3H), 1.37 (d, J=7.2 Hz, 3H). Solutionof amine 4.2(2), (S)-4.2(2) or (R)-4.2(2) (110 mg, 0.46 mmol) and4-isothiocyanato-2-(trifluoromethyl)-benzonitrile 3.2 (144 mg, 0.55mmol) in DMF (2 ml) was stirred at 90° C. for 12 h in microwave oven,then additional portion of4-isothiocyanato-2-(trifluoromethyl)-benzonitrile 3.2 (50 mg, 0.19 mmol)was added and stirring was continued for another 12 h. The reactionmixture was evaporated in vacuo, and by HPLC methodN-methyl-4-{5-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl}-2-fluorobenzamide1.2(2), orN-methyl-4-{(S)-5-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl}-2-fluorobenzamide(S)-1.2(2) orN-methyl-4-{(R)-5-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl}-2-fluorobenzamide(R)-1.2(2) were isolated, respectively. The apparent inhibition constantof androgen receptors (K_(i))for these compounds are: K_(i)^(1.2(2))=140.2 nM, K_(i) ^((S)-1.2(2))=106.7 nM

K_(i) ^((R)-1.2(2))=73.6 nM, respectively. LCMS (M+H)⁺ 451. ¹H NMR(CDCl₃, 400 MHz): 8.28 (t, J=8.6 Hz, 1H), 8.01 (d, J=8.0 Hz, 1H), 7.94(d, J=1.2 Hz, 1H), 7.81 (dd, J₁=8.0 Hz, J₂=1.2 Hz, 1H), 7.48 (dd,J₁=12.4 Hz, J₂=1.6 Hz, 1H), 7.36 (dd, J₁=8.4 Hz, J₂=1.6 Hz, 1H), 6.72(br. m, 1H), 4.83 (q, J=7.2 Hz, 1H), 3.08 (d, J=4.8 Hz, 3H), 1.60 (d,J=7.2 Hz, 3H).

EXAMPLE 3

Synthesis ofN-methyl-4-{2-thio-3-[3-(trifluoromethyl)-4-cyanophenyl]-hydantoin-1-yl}-2-fluorobenzamides1.2.2 and 1.2.3 (general method). Solution of the correspondingN-methyl-2-fluoro-4-[(1-cyanomethyl)amino]benzamide 4.1 (0.75 mmol) and4-isothiocyanato-2-(trifluoromethyl)benzonitrile 3.2 (342 mg, 1.5 mmol)in DMF (3 ml) was stirred at 110° C. for 12 h in microwave oven. Thereaction mixture was dissolved in MeOH (30 ml), 1N HCl (7.5 ml) wasadded and the resultant mixture was boiled for 1.5 h. The solution wasevaporated in vacuo, treated with water, the solid was filtered off,washed with water and dried in vacuo. The product was isolated by HPLCmethod. It gave:N-methyl-4-[5-methyl-5-(methoxymethyl)-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]-imidazolidin-1-yl]-2-fluorobenzamide1.2.2(1), K_(i) ^(1.2.2(1))=115.9 nM, which was separated to enantiomersby means of high pressure liquid chromatography on Chiralpak HD-H 25×1cm (Chiral Technologies Inc., USA). Mixture of 80% n-hexane, 20%2-propanol and 0.02% triethylamine was used as eluent. Flowrate was 4ml/min. It gave optically pure isomers (R)-1.2.2(1) and (S)-1.2.2(1),K_(i) ^((R)-1.2.2(1))=53.3 nM, K_(i) ^((S)-1.2.2(1))=721.5 nM. LCMS(M+H)⁺ 495. ¹H NMR (CDCl₃, 400 MHz): 8.28 (t, J=8.4 Hz, 1H), 7.99 (d,J=8.0 Hz, 1H), 7.92 (s, 1H), 7.80 (d, J=8.0 Hz, 1H), 7.29 (dd, J₁=8.8Hz, J₂=1.2 Hz, 1H), 7.21 (dd, J₁=11.6 Hz, J₂=1.2 Hz, 1H), 6.72 (q, J=4.4Hz, 1H), 3.71 (d, J=10.0 Hz, 1H), 3.43 (s, 3H), 3.35 (d, J=10.0 Hz, 1H),3.09 (d, J=4.4 Hz, 3H), 1.52 (s, 3H);

N-methyl-4-{5-[(benzyloxy)methyl]-5-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl}-2-fluorobenzamide1.2.2(2). LCMS (M+H)⁺ 571. ¹H NMR (CDCl₃, 400 MHz): 8.22 (t, J=8.4 Hz,1H), 7.96 (d, J=8.0 Hz, 1H), 7.86 (s, 1H), 7.70 (dd, J₁=8.0 Hz, J₂=1.2Hz, 1H), 7.39 (m, 3H), 7.29 (m, 2H), 7.25 (dd, J₁=8.4 Hz, J₂=1.6 Hz,1H), 7.18 (dd, J₁=8.4 Hz, J₂=1.6 Hz, 1H), 6.71 (q, J=4.8 Hz, 1H), 4.59(m, 2H), 3.79 (d, J=10.2 Hz, 1H), 3.45 (d, J=10.2 Hz, 1H), 3.08 (d,J=4.8 Hz, 3H), 1.51 (s, 3H);

ethyl{4-methyl-3-(4-methylcarbamoyl-3-fluorophenyl)-5-oxo-2-thioxo-1-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-4-yl}acetate1.2.2(4) (R1=CH₃, R4=CH₃, R5=CH₂COOC₂H₅). LCMS (M+H)⁺ 536. ¹H NMR(CDCl₃, 400 MHz): 8.26 (t, J=8.4 Hz, 1H), 8.01 (d, J=8.0 Hz, 1H), 8.00(s, 1H), 7.90 (dd, J₁=8.0 Hz, J₂=1.6 Hz, 1H), 7.18 (dd, J₁=8.0 Hz,J₂=1.6 Hz, 1H), 7.10 (dd, J₁=8.0 Hz, J₂=1.6 Hz, 1H), 6.78 (q, J=4.8 Hz,1H), 4.26 (m, 1H), 3.13 (d, J=18.0 Hz, 1H), 3.09 (d, J=4.8 Hz, 3H), 2.64(d, J=18.0 Hz, 1H), 1.67 (s, 3H), 1.31 (t, J=7.0 Hz, 3H);

N-methyl-4-{4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]-7-oxa-1,3-diazaspiro[4.4]non-1-yl}-2-fluorobenzamide1.2.3(1), K_(i) ^(1.2.3(1))=33.9 nM. LCMS (M+H)⁺ 493. ¹H NMR (CDCl₃, 400MHz): 8.30 (t, J=8.4 Hz, 1H), 8.02 (d, J=8.4 Hz, 1H), 7.98 (d, J=1.6 Hz,1H), 7.85 (dd, J₁=8.4 Hz, J₂=1.6 Hz, 1H), 7.34 (dd, J₁=8.4 Hz, J₂=1.6Hz, 1H), 7.25 (dd, J₁=11.8 Hz, J₂=1.6 Hz, 1H), 6.78 (q, J=4.4 Hz, 1H),4.43 (d, J=10.0 Hz, 1H), 4.16 (d, J=10.0 Hz, 1H), 3.96 (m, 1H), 3.75 (m,1H), 3.09 (d, J=4.4 Hz, 3H), 2.74 (m, 1H), 2.48 (m, 1H);

N-methyl-4-{4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]-8-oxa-1,3-diazaspiro[4.5]dec-1-yl}-2-fluorobenzamide1.2.3(2). LCMS (M+H)⁻ 507. ¹H NMR (CDCl₃, 400 MHz): 8.32 (t, J=8.4 Hz,1H), 8.01 (d, J=8.0 Hz, 1H), 7.95 (s, 1H), 7.83 (d, J=8.0 Hz, 1H), 7.20(d, J=8.4 Hz, 1H), 7.10 (d, J=8.0 Hz, 1H), 6.73 (br. m, 1H), 4.18 (m,2H), 3.94 (m, 2H), 3.09 (d, J=4.4 Hz, 3H), 2.07 (m, 4H);

N-methyl-4-{8-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]-1,3,8-triazaspiro[4.5]dec-1-yl}-2-fluorobenzamide1.2.3(3). K_(i) ^(1.2.3(3))=39.2 nM, IC₅₀=170 nM. LCMS (M+H)⁺ 520. ¹HNMR (DMSO-d₆, 400 MHz): 10.09 (br. s, 1H), 8.48 (q, J=4.4 Hz, 1H), 8.43(d, J=8.4 Hz, 1H), 8.29 (s, 1H), 8.11 (d, J=8.4 Hz, 1H), 7.84 (t, J=8.0Hz, 1H), 7.42 (d, J=10.4 Hz, 1H), 7.30 (d, J=8.0 Hz, 1H), 3.50 (m, 4H),2.80 (d, J=4.4 Hz, 3H), 2.78 (s, 3H), 2.72 (d, J=14.0 Hz, 1H), 2.16 (m,2H).

Such salts of the compounds of general formula 1.2.3 of the presentinvention as hydrochloride, hydrobromide, phosphate, nitrate,perchlorate, sulfate, acetate could be formed by methods well known inthe art. For example, the compound 1.2.3(3) was dissolved indichloromethane and a saturated solution of HCl in dioxane was added.TheN-methyl-4-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]-1,3,8-triazaspiro[4.5]dec-1-yl}-2-fluorobenzamidehydrochloride salt 1.2.3(3)*HCl obtained in the precipitate was washedwith dioxane, evaporated and dried. LCMS (M+H)⁺ 520.

EXAMPLE 4

Synthesis ofN-methyl-4-[5-(hydroxymethyl)-5-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl]-2-fluorobenzamide1.2.2(3). BBr₃ (53 mkl, 0.55 mmol) was added dropwise to solution ofN-methyl-4-[5-methyl-5-(methoxymethyl)-4-oxo-2-thioxo-3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1yl]-2-fluorobenzamide(55 mg, 0.11 mmol) in CH₂Cl₂ (1.5 ml) in argon atmosphere at −78° C. Thereaction mixture was stirred at −78° C. for 3 h and then for another 3 hat room temperature. After the reaction was completed the excess of BBr₃was neutralized by addition of 5% Na₂CO₃ solution (10 ml), the productwas extracted with AcOEt, dried over Na₂SO₄, evaporated in vacuo, and byHPLC methodN-methyl-4-[5-(hydroxymethyl)-5-methyl-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-1-yl]-2-fluorobenzamide1.2.2(3) was isolated, K_(i) ^(1.2.2(3))=46.3 nM, ¹H NMR (DMSO-d₆, 400MHz): 8.43 (br. m, 1H), 8.39 (d, J=8.4 Hz, 1H), 8.13 (s, 1H), 7.98 (d,J=8.4 Hz, 1H), 7.78 (t, J=8.0 Hz, 1H), 7.42 (d, J=10.8 Hz, 1H), 7.37 (d,J=8.0 Hz, 1H), 5.93 (t, J=4.4 Hz, 1H), 3.81 (dd, J₁=11.6 Hz, J₂=4.4 Hz,1H), 3.45 (dd, J₁=11.6 Hz, J₂=5.0 Hz, 1H), 2.79 (d, J=4.0 Hz, 3H), 1.38(s, 3H).

EXAMPLE 5

Synthesis of{4-methyl-3-(4-methylcarbamoyl-3-fluorophenyl)-5-oxo-2-thioxo-1-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-4-yl}aceticacid 1.2.2(5) (R1=CH₃, R4=CH₃, R5=CH₂COOH). Solution of NaOH (7 mg,0.172 mmol) in water (0.5 ml) was added to the solution of ester (46 mg,0.086 mmol) 1.2.2(4) in alcohol (2 ml), and the reaction mixture wasstirred for 12 h (LCMS control). The solvent was evaporated, isopropanol(2 ml) and HCl (15 mkl, 0.172 mmol) were added, filtered and evaporatedagain in vacuo.{4-Methyl-3-(4-methylcarbamoyl-3-fluorophenyl)-5-oxo-2-thioxo-1-[3-(trifluoromethyl)-4-cyanophenyl]imidazolidin-4-yl}aceticacid 1.2.2(5) was isolated by HPLC method. LCMS (M+H)⁺ 469. ¹H NMR(DMSO-d₆, 400 MHz): 13.31 (br. s, 1H), 8.44 (m, 2H), 8.10 (s, 1H), 7.95(d, J=7.6 Hz, 1H), 7.81 (t, J=8.0 Hz, 1H), 7.25 (d, J=10.8 Hz, 1H), 7.19(d, J=8.0 Hz, 1H), 3.16 (d, J=17.6 Hz, 1H), 2.79 (d, J=3.6 Hz, 3H), 2.70(d, J=17.6 Hz, 1H), 1.59 (s, 3H).

EXAMPLE 6

Synthesis of4-[3-[3-(trifluoromethyl)-4-cyanophenyl]-2-oxo-tetrahydro-pyrimidin-1(2H)-yl]-N-methyl-2-fluorobenzamide1.3.1. K₂CO₃ (109 mg, 0.79 mmol) and 1,3-dibromopropane (32 mkl, 0.32mmol) were added to a solution of4-[4-cyano-3-(trifluoromethyl)-phenylcarbamoylamino]-N-methyl-2-fluorobenzamide(100 mg, 0.26 mmol) 2 in DMF (2 ml). Mixture was stirred at 90° C. In 18h another portion of K₂CO₃ (109 mg) and 1,3-dibromopropane (32 mkl) wereadded and stirring was continued at the same temperature. Addition wasrepeated by 2 more times. After the reaction was completed the mixturewas evaporated in vacuo, the residue was dissolved in chloroform, washedwith water, dried over Na₂SO₄, the solvent was evaporated. The productwas isolated by colomn chromatography on SiO₂ (eluent—AcOEt). LCMS(M+H)⁺ 421. ¹H NMR (DMSO-d₆, 400 MHz): 8.17 (br. m, 1H), 8.13 (d, J=8.4Hz, 1H), 8.07 (d, J=2.0 Hz, 1H), 7.83 (dd, J₁=8.4 Hz, J₂=2.0 Hz, 1H),7.62 (t, J=8.4 Hz, 1H), 7.38 (dd, J₁=12.4 Hz, J₂=2.0 Hz, 1H), 7.29 (dd,J₁=8.4 Hz, J₂=2.0 Hz, 1H), 3.90 (t, J=5.8 Hz, 2H), 3.81 (t, J=5.8 Hz,2H), 2.77 (d, J=4.8 Hz, 3H), 2.21 (m, 2H).

EXAMPLE 7

Synthesis ofN-methyl-4-{[3-(trifluoromethyl)-4-cyanophenyl]-2,4-dioxo-tetrahydropyrimidin-1(2H)-yl}benzamide1.3.2(1), (X═O, R1=CH₃). Ethyl acrylate (8 g, 80 mmol) and DBU (0.81 g,5.4 mmol) were added to a solution of 4-amino-N-methyl-2-fluorobenzamide(9 g, 53.6 mmol) in DMSO (90 ml) and stirring was continued for 24 h at70° C. (LCMS control). The reaction mixture was subjected tolyophilization, the residue was recrystallized from aqueous alcohol. Itgave ethyl N-[4-(methylcarbamoyl)-3-fluorophenyl]-β-alaninate 5. LCMS(M+H)⁻ 269. ¹H NMR (DMSO-d₆, 400 MHz) δ 7.57 (br. s, 1H), 7.48 (t, J=8.8Hz, 1H), 6.47 (br. s, 1H), 6.42 (d, J=8.8 Hz, 1H), 6.33 (d, J=14.8 Hz,1H), 4.07 (q, J=7.2 Hz, 2H), 3.32 (br. m, 2H), 2.73 (d, J=4.4 Hz, 3H),2.55 (t, J=6.4 Hz, 2H), 1.18 (t, J=7.2 Hz, 3H). The solution of4-isocyanato-2-(trifluoromethyl)benzonitrile (425 mg, 1.87 mmol) 3.1 andethyl N-[4-(methylcarbamoyl)-3-fluorophenyl]-β-alaninate (500 mg, 1.87mmol) 5 (R1=CH₃) in CH₂Cl₂ (10 ml) was stirred for 5 h. The reactionmixture was evaporated in vacuo, and the product was isolated by colomnchromatography on SiO₂ (eluent—n-hexane:AcOEt:Et₃N=1:1:0.03). It gaveethylN-[4-(methylcabamoyl)-3-fluorophenyl]-N-{[3-(trifluoromethyl)-4-cyanophenyl]-carbamoyl}-β-alaninate6(1) (R1=CH₃, X═O). LCMS (M+H)⁺ 481. HCl (2.5 ml) was added to thesolution of ethylN-[4-(methylcabamoyl)-3-fluorophenyl]-N-{[3-(trifluoromethyl)-4-cyanophenyl]-carbamoyl}-β-alaninate(500 mg, 1.04 mmol) 6(1) in AcOH (5 ml) and the resultant mixture wasstirred for 15 h. The reaction mixture was poured into water, theproduct was extracted with EtOAc. Organic layer was dried over Na₂SO₄,evaporated in vacuo and by means of colomn chromatography on SiO₂(eluent—n-hexane:AcOEt=1:1)N-methyl-4-{[3-(trifluoromethyl)-4-cyanophenyl]-2,4-dioxo-tetrahydropyrimidin-1(2H)-yl}benzamide1.3.2, (X═O, R1=CH₃) was isolated; K_(i) ^(1.3.2(1))=85.6 nM, LCMS(M+H)⁺ 435. ¹H NMR (DMSO-d₆, 400 MHz): δ 8.29 (d, J=7.6 Hz, 1H), 8.23(br. m, 1H), 8.11 (s, 1H), 7.89 (d, J=2.0 Hz, 1H), 7.67 (t, J=8.4 Hz,1H), 7.39 (d, J=12.4 Hz, 1H), 7.33 (d, J=8.4 Hz, 1H), 4.02 (t, J=6.4 Hz,2H), 3.03 (t, J=6.4 Hz, 2H), 2.77 (d, J=4.4 Hz, 3H).

EXAMPLE 8

Synthesis ofN-methyl-4-{[3-(trifluoromethyl)-4-cyanophenyl]-4-oxo-2-thioxo-tetrahydropyrimidin-1(2H)-yl}benzamide1.3.2(2), (X=5, R1=CH₃). A solution of4-isothiocyanato-2-(trifluoromethyl)benzonitrile (320 mg, 1.51 mmol) 3.2and ethyl N-[4-(methylcarbamoyl)-3-fluorophenyl]-β-alaninate (404 mg,1.51 mmol) 5 (R1=CH₃) in DMF (8 ml) was heated at 60° C. in microwavestove for 8 h. The reaction mixture was evaporated in vacuo, and bymeans of colomn chromatography on SiO₂ (eluent—n-hexane:AcOEt=1:2) ethylN-[4-(methylcarbamoyl)-3-fluorophenyl]-N-{[3-(trifluoromethyl)-4-cyanophenyl]thiocarbamoyl}-β-alaninate6(2) (R1=CH₃, X═S) was isolated. LCMS (M+H)⁺ 497. A solution of NaOH (32mg, 0.8 mmol) in water (0.25 ml) was added to a solution of ester (200mg, 0.4 mmol) 6(2) in alcohol (1 ml), and the resultant mixture wasstirred at 80° C. for 2 h (LCMS control), cooled and neutralized withHCl (69 mkl, 0.8 mmol), evaporated in vacuo, the residue was extractedwith hot isopropanol and evaporated in vacuo again. It gaveN-[4-(methylcarbamoyl)-3-fluorophenyl]-N-{[3-(trifluoromethyl)-4-cyanophenyl]thiocarbamoyl}-β-alanine6(3) (R1=CH₃, X═S). LCMS (M+H)⁺ 469. TBTU (86 mg, 0.36 mmol) anddiisopropylethylamine (110 mg, 0.84 mmol) were added to the solution ofthe prepared acid (114 mg, 0.24 mmol) 6(3) in DMF (1.5 ml). The reactionmixture was stirred at 45° C. for 15 h. When the reaction was completed(LCMS control) the solution was poured into water and extracted withEtOAc. The organic layer was dried over Na₂SO₄, evaporated in vacuo andby HPLC methodN-methyl-4-{[3-(trifluoromethyl)-4-cyanophenyl]-4-oxo-2-thioxo-tetrahydropyrimidin-1(2H)-yl}-2-fluorobenzamide1.3.2(2) (R1=CH₃, X═S) was isolated; K_(i) ^(1.3.2(2))=95.2 nM. LCMS(M+H)⁺ 451. ¹H NMR (DMSO-d₆, 400 MHz): 8.35 (q, J=4.4 Hz, 1H), 8.27 (d,J=8.0 Hz, 1H), 8.06 (d, J=1.6 Hz, 1H), 7.83 (dd, J₁=8.0 Hz, J₂=1.6 Hz,1H), 7.71 (t, J=8.2 Hz, 1H), 7.42 (dd, J₁=11.0 Hz, J₂=1.8 Hz, 1H), 7.33(dd, J₁=8.2 Hz, J₂=1.8 Hz, 1H), 4.13 (t, J=6.8 Hz, 2H), 3.17 (t, J=6.8Hz, 2H), 2.78 (d, J=4.4 Hz, 3H).

EXAMPLE 9

Synthesis ofN-methyl-2-fluoro-4-[4-[3-(trifluoromethyl)-4-cyanophenyl]-3,5-dioxo-1,2,4-triazolidin-1-yl]benzamide1.4 (R1=CH₃). 2.5M Solution of NaNO₂ (2.38 ml) was added dropwise to asolution of 4-amino-N-methyl-2-fluorobenzamide (1 g, 5.95 mmol) in B 5NHCl (3.1 ml), so that the temperature of the reaction mixture did notexceed 5° C. The mixture was stirred for additional 30 min at the sametemperature, after that the prepared solution was added drop by drop toa suspension of SnCl₂*2H₂O (4.03 g, 17.9 mmol) in HCl (4.2 ml) at 0° C.,and stirring was continued for 2 h at the same temperature. Precipitatedsolid was filtered off, dissolved in water (40 ml) and NaOH was added tostrongly basic reaction. The mixture was extracted with ether (3*100ml), dried over MgSO₄ and evaporated in vacuo. It gave4-hydrazino-N-methyl-2-fluorobenzamide 7 (R1=CH₃). LCMS (M+H)⁺ 184. ¹HNMR (CDCl₃, 400 MHz): 7.96 (t, J=8.4 Hz, 1H), 6.64 (br. m, 1H), 6.60 (t,J=1.6 Hz, 1H), 6.57 (dd, J₁=7.2 Hz, J₂=2.0 Hz, 1H), 5.60 (br. s, 1H),3.66 (br. s, 2H), 3.00 (dd, J₁=4.8 Hz, J₂=1.2 Hz, 1H). The solution of4-isocyanato-2-(trifluoromethyl)benzonitrile (59 mg, 0.27 mmol) 3.1 indioxane (2 ml) was added to a solution of4-hydrazino-N-methyl-2-fluorobenzamide (54 mg, 0.29 mmol) 7 in dioxane(3 ml), and the resultant mixture was stirred for 2 h. Then dioxane wasdistilled in vacuo, the residue was crumbled with ether, filtered offand dried in vacuo. It gave2-[(4-methylcarbamoyl)-3-fluorophenyl]-N-[3-(trifluoromethyl)-4-cyanophenyl]-hydrazinecarboxamide 8(1) (R1=CH₃). LCMS (M+H)⁺ 405. ¹H NMR (DMSO-d₆, 400 MHz):9.65 (br. s, 1H), 8.72 (br. s, 1H), 8.37 (s, 1H), 8.25 (br. s, 1H), 8.03(br. m, 1H), 7.88 (d, J=8.8 Hz, 1H), 7.58 (m, 2H), 6.63 (d, J=8.4 Hz,1H), 6.48 (d, J=14.0 Hz, 1H), 2.77 (d, J=4.4 Hz, 3H). Triethylamine (56mkl, 0.4 mmol) and diphosgene (27 mkl, 0.22 mmol) were added one afteranother to2-[(4-methylcarbamoyl)-3-fluorophenyl]-N-[3-(trifluoromethyl)-4-cyanophenyl]-hydrazinecarboxamide (80 mg, 0.2 mmol) 8(1) in dichloroethane (2 ml). Thereaction mixture was stirred in a closed vial at 80° C. for 15 h. Thesolvent was evaporated in vacuo and the residue was subjected tochromatography on SiO₂ (eluent—CH₂Cl₂:MeOH, gradient from 100:1 till20:1). It gaveN-methyl-2-fluoro-4-[4-[3-(trifluoromethyl)-4-cyanophenyl]-3,5-dioxo-1,2,4-triazolidin-1-yl]benzamide1.4 (R1=CH₃). K_(i) ^(1.4)=55.2 nM, LCMS (M+H)⁺ 422. ¹H NMR (DMSO-d₆,400 MHz): 11.53 (s, 1H), 8.22 (br. m, 1H), 8.16 (d, J=8.8 Hz, 1H), 7.99(dd, J₁=8.8 Hz, J₂=1.6 Hz, 1H), 7.95 (d, J=1.6 Hz, 1H), 7.81 (t, J=8.4Hz, 1H), 7.69 (dd, J₁=8.8 Hz, J₂=2.0 Hz, 1H), 7.62 (dd, J=12.0 Hz,J₂=2.0 Hz, 1H), 2.79 (d, J=4.4 Hz, 3H).

EXAMPLE 10

Determination of antagonistic activity of cyclic N,N′-diarylthioureasand N,N′-diarylureas of the general formula 1 and their analog MDV3100towards androgen receptors. The ability of novel cyclicN,N′-diarylthioureas and N,N′-diarylureas of the general formula 1 andMDV3100 agent to block androgen receptors was determined by theireffectiveness of inhibition of dihydrotestosterone stimulated expressionof prostate specific antigen (PSA) in cancer cells of human prostrateLNCap, derived from the American Tissue Culture Collection (ATCC, USA).These cells are sensitive towards 5-a-dihydrotestosterone (DHT) and inits presence produce cancer markers (PSA). The cells were cultured inRPMI 1640 medium (Invitrogen, USA) containing 10% calf serum (Hyclone,USA), 1% antibacterial/antifungal mixture (Sigma, USA) and 4,5% glucose.Before the experiment the cells were washed and suspended in the samemedium in which, however, instead of calf serum the serum which had beentreated with charcoal for removal of hormone traces was used. The cellswere embedded into wells of 96-well plates by 100 μl per cell (10 000cells) and left for 4 days in incubator at 37° C. (100% humidity) inatmosphere of 95% air/5% CO₂. After incubation cyclicN,N′-diarylthioureas or N,N′-diarylureas of the general formula 1 wereadded to the cells in various concentrations, and then −20 nM DHT(concentration corresponding to 80-90% of maximum stimulation). Thecells were left for 5 days for additional incubation under the sameconditions. After that the samples of supracellular medium were taken onanalysis for PSA content. The test was carried out according to theprotocol, recommended by manufacturer of the kit for determination ofPSA (Alpha Diagnostic International, USA). After wetting the wellscontaining PSA antibodies attached to their bottom to each well 25 μl ofthe tested compounds and 100 μl of PSA antibodies conjugated previouslywith horseradish peroxidase were added successively.

After incubation at room temperature for 30 minutes, the contents of thewells were removed, the wells were washed several times, and then 100 μlof chromogenic substrate of peroxidase was added to each well. Plateswere held for 15 min. at room temperature, after that 50 μl of stopsolution was added to every well; at that a dye is formed the absorptionintensity of which was measured at 450 nM; the value obtained isproportional to PSA concentration in the sample. Based on the dependenceof lowering of PSA synthesis, caused by dihydrotestosterone (DHT), onthe concentration of the tested compounds, dose-response curves wereplotted, from which IC₅₀ values were determined. They were used forcalculation of the values of apparent inhibition constants (K_(i)) forthe compounds of the general formula I according to Cheng-Prusoffequation. [Cheng, Y., Prusoff, W. H. “Relationship between theinhibition constant (K_(i))and the concentration of inhibitor whichcauses 50 per cent inhibition (IC₅₀) of an enzymatic reaction”. BiochemPharmacol. (1973) 22 , 3099-3108]:

K _(i)=IC₅₀/(1+L/K _(D)),

wherein L—agonist concentration (DHT), K_(D)—receptor activationconstant, numerically equal to EC₅₀ value, which is determined in everyexperiment according to dependence of stimulation of PSA synthesis onDHT concentration.

The data obtained given in the corresponding examples testify that novelandrogen receptor antagonists, in some cases, are more active thanMDV3100, tested under the same conditions as a compound for comparison,for which K_(i) ^(MDV3100)=79.5 nM.

EXAMPLE 11

Determination of maximum tolerated dose of novel antagonists 1.2.2(1),and 1.2.3(3) and its analog MDV3100. Maximum tolerated dose (MTD) ofnovel antagonists 1.2.2(1) and 1.2.3(3) and its analog MDV3100 weredetermined in experiments on male mice of CD1 line at peroraladministration 1 time a day within 5 days in doses 10, 30 and 100 mg/kg.The compound was dissolved in sterile water with addition of Twin-80.Sterile water with Twin-80 was introduced to control animals (Placebogroup). Body weight was appreciated, and also animals' mortality rate.Statistical comparison of groups was carried out according tonon-parametric test ANOVA, with the use of Statistica programme.

At the administering compounds 1.2.2(1) or 1.2.3(3) in dose up to 100mg/kg mice death was not observed. On the 3rd-4th day body weight ofmice in the group, received the tested compound in dose 100 mg/kg wasless in comparison with the body weight of control animals, however,statistical significance at this was not observed (FIG. 1). The datashow that compound 1.2.2(1) and 1.2.3(3) has MTD>100 mg/kg.

At the administering MDV3100 in doses 10 and 30 mg/kg mice death was notobserved. In the group of mice to which the tested compound wasintroduced in dose 100 mg/kg, the body weight began to lower on the3^(rd) day. On the 5^(th) day body weight of this group of animalsstatistically differed from body weight of animals from Placebo group(p=0.002, FIG. 2). One animal died. The data show that compound MDV3100has MTD˜30 mg/kg.).

EXAMPLE 12

Preparation of medicament in the form of tablets. Starch (1600 mg),grained lactose (1600 mg), talcum (400 mg) andN-methyl-4-[5-methyl-5-(methoxymethyl)-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]-imidazolidin-1-yl]-2-fluorobenzamide(R)-1.2.2(1) (1000 mg) mixed together and pressed in a brick. Preparedbrick was crushed to granules and riddled through sieves, gatheringgranules of 14-16 mesh size. The obtained granules were pelletised intablets of suitable form of 560 mg by weight each.

EXAMPLE 13

Preparation of medicament in the form of capsules.N-Methyl-4-[5-methyl-5-(methoxymethyl)-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]-imidazolidin-1-yl]-2-fluorobenzamide(R)-1.2.2(1) was carefully mixed with lactose powder in ratio 2:1. Theprepared pharmaceutical composition was packed on 300 mg into gelatinouscapsules of suitable size.

EXAMPLE 16

Preparation of medicament in the form of compositions for intramuscular,intraperitoneal or hypodermic injections.N-Methyl-4-[5-methyl-5-(methoxymethyl)-4-oxo-2-thioxo-3-[3-(trifluoromethyl)-4-cyanophenyl]-imidazolidin-1-yl]-2-fluorobenzamide(R)-1.2.2(1) (500 mg) was dissolved in the mixture of chlorobutanole(300 mg), propylene glycol (2 ml), and water for injections (100 ml).The prepared solution was filtered and placed in 1 ml ampoules whichwere sealed up and sterilized in an autoclave.

INDUSTRIAL APPLICABILITY

The present invention could be used in medicine, veterinary,biochemistry.

1. A cyclic N,N′-diarylthioureas and N,N′-diarylurea compound of general formula 1, or an optical (R)- or (S)-isomer, or a pharmaceutically acceptable salt thereof, exhibiting properties of androgen receptor antagonist,

wherein: X represents oxygen or sulfur; m=0 or 1; R1 represents C₁-C₃alkyl; R2 and R3 represent hydrogen; or R2 and R3 together with the carbon atom they are attached to form C═O group; R4 and R5 represent hydrogen; or R4 represents hydrogen, R5 represents methyl; or R4 represents methyl, R5 represents CH₂R6 group in which R6 is C₁-C₃ alkoxycarbonyl, carboxyl, hydroxyl group optionally substituted with methyl or benzyl; or R4 and R5 together with the carbon atom they are attached to form 5- or 6-membered saturated heterocycle including at least one oxygen atom or nitrogen atom optionally substituted with methyl, or R4 and R5 together with the carbon atom they are attached to represent NH-group.
 2. The compound of claim 1 selected from cyclic N,N′-diarylthioureas and N,N′-diarylureas of general formulas 1.2, 1.3 or 1.4, or an optical (R)- or (S)-isomer, or a pharmaceutically acceptable salt thereof,


3. The compound of claim 2 selected from the group consisting of cyclic N,N′-diarylthioureas of formulas 1.2(1), 1.2(2), 1.2.2, 1.2.3, optical (R)-isomers (R)-1.2(2), (R)-1.2.2, (R)-1.2.3, optical (S)-isomers (S)-1.2(2), (S)-1.2.2, and (S)-1.2.3, or a pharmaceutically acceptable salt thereof,

wherein: R4 and R5 together with the carbon atom they are attached to form 5- or 6-membered saturated heterocycle comprising at least one oxygen atom or nitrogen atom optionally substituted with methyl; R6 has the above meanings
 4. The compound of claim 2 selected from the group consisting of compounds of formulas 1.2.2(1), 1.2.2(2), 1.2.2(3), 1.2.3(1), 1.2.3(2), 1.2.3(3), optical (R)-isomers (R)-1.2.2(1), (R)-1.2.2(2), (R)-1.2.2(3), (R)-1.2.3(1), optical (S)-isomers (S)-1.2.2(1), (S)-1.2.2(2), (S)-1.2.2(3), and (S)-1.2.3(1), or a pharmaceutically acceptable salt thereof,

wherein R6 is hydroxyl group optionally substituted with methyl or benzyl.
 5. A process for the preparation of a compound of formula 1.2, or an optical (R)- or (S)-isomer in any of claims 1-4 by mixing a corresponding 4-(cyanomethyl)amino-benzamide compound of formula 4.1 or a (4-carbamoyl-phenylamino)-acetic acid of formula 4.2, or an optical (R)- and (S)-isomer with an isothiocyanate of formula 3.2,


6. An anticancer agent comprising at least one cyclic N,N′-diarylthiourea or N,N′-diarylurea compound of the general formula 1 according to any of claims 1-4 exhibiting properties of androgen receptor antagonists.
 7. A pharmaceutical composition exhibiting properties of androgen receptor antagonist comprising as an active component an anticancer agent according to claim 6 and at least one pharmaceutically acceptable carrier, inert excipient or solvent.
 8. A method for preparation of the pharmaceutical composition according to claim 7 by mixing an anticancer agent according to claim 6 with inert filler and/or solvent.
 9. A medicament in the form of a tablet, sheath or an injection for treating cancerous disease comprising an anticancer agent according to claim 6 or a pharmaceutical composition according to claim
 7. 10. The medicament according to claim 9 intended for prostate cancer treatment.
 11. A method for treating a cancerous disease, including prostate cancer, comprising administering a medicament according to any of claims 9-10, or a pharmaceutical composition according to claim 7, or an anticancer agent according to claim 6 to a subject in need thereof.
 12. An androgen receptor antagonist according to claim 1 for investigation of molecular mechanism of inhibition and activation of androgen receptors. 